产品&服务

Cellufine™ 层析介质

纤维素珠体层析介质

Cellufine™ 层析介质基于球形纤维素珠体,具有高化学稳定性、高机械强度和固有的生物兼容性。Cellufine™ 产品适用于所有层析技术。凝胶过滤、离子交换、亲和性和疏水作用层析介质可用于制备各种目标分子。

应用

层析介质产品范围

Typical Product Range
TypeApplicationsSpecification
Ion Exchange
Cellufine™ IEX
For initial separation and final polishing of proteins, peptides and enzymes by electrostatic interaction Ion exchange resins (DEAE, QA, CM, S)
MAX IEX (High performance Ion exchange)
Graft polymer modification
Cellufine™ MAX GS
Mix mode
Cellufine™ MAX IB
For high performance impurities removal from target molecules Strong cation exchange on Graft chain
Mix mode Anion exchange resin
Affinity
Cellufine™ Sulfate
Cellufine™ ET Clean
Cellufine™ Chlete, etc.
For high specificity separation of a wide range of molecules by specific interaction Heparin mimic for virus & heparin binding proteins
Endotoxin removal
IMAC metal binding molecules
Molecules with diol & EPO
Nucleic acid related molecules
Hydrophobic Interaction
Cellufine™ Butyl
Cellufine™ Phenyl
For purification of proteins and enzymes by hydrophobic interaction Butyl
Phenyl
Gel filtration
Cellufine™ GCL-2000HF
Cellufine™ GH-25
For purification of bio-molecules and proteins by molecular size.
For salt and solvent removal and buffer exchange
MW 50 - 3000 kD
3 kD gel exclusion limit
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离子交换

Cellufine™ IEX 系列配体的化学结构

▶Strong Anion Exchange (Q) Weak Anion Exchange (DEAE)
Cellufine™ MAX Q-h
Cellufine™ MAX Q-r
Cellufine™ Q-500
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Cellufine™ MAX DEAE
Cellufine™ A-200
Cellufine™ A-500
Cellufine™ A-800
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Strong Cation Exchange (S) Weak Cation Exchange (CM)
Cellufine™ MAX S-h
Cellufine™ MAX S-r
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Cellufine™ MAX CM
Cellufine™ C-500
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在高流速下优越的结合能力

即使在高流速下,Cellufine™ MAX IEX 介质与其他琼脂糖或丙烯酸介质相比,其DBC较高。

Figure 4
BSA DBC at 10% Breakthrough for Various Flow Rates

▶Strong Anion Media ▶Weak Anion Media
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BSA concentration: 1 mg/mL
Column: 5 mm I.D. x 100 mm
Buffer: 50 mM Tris-HCl (pH 8.5)

BSA concentration: 1 mg/mL
Column: 5 mm I.D. x 50 mm
Buffer: 50 mM Tris-HCl (pH 8.5)

IgG DBC at 10% Breakthrough for Various Flow Rates

▶Strong Cation Media ▶Weak Cation Media
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IgG concentration: 1 mg/mL (polyclonal, human)
Column: 5 mm I.D. x 100 mm
Buffer: 10 mM Na-acetate, 50 mM NaCl (pH 4.3)

IgG concentration: 1 mg/mL (polyclonal, human)
Column: 5 mm I.D. x 50 mm
Buffer: 10 mM Na-acetate (pH 5.6)

高耐压性实现高效纯化

Cellufine™ MAX IEX 层析介质可在高压条件下有效运行,从而实现生物技术药品的高效纯化。
右图显示了用于大柱中Cellufine™ MAX IEX 系列介质的压力流量曲线。所有Cellufine™ MAX IEX 系列介质够都可以在高压下运行。
Pressure Flow Curves of Cellufine™ MAX IEX Chromatography Media
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Column: 300 mm I.D. x 200 mm
Mobile phase: pure water (20 ºC)

优越的吸附和分离度

Cellufine™ MAX IEX 层析介质针对高吸附和分离度进行了优化。下图显示了模型蛋白质的分离。观察到良好的分离曲线。

Separation of Model Proteins by Cellufine™ MAX Q-h and Cellufine™ MAX DEAE Separation of Model Proteins by Cellufine™ MAX S-h and Cellufine™ MAX CM
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Column: 6.6 mm I.D. x 50 mm
Equilibration buffer (A): 50 mM Tris-HCl (pH 8.5)
Elution buffer (B): 1 M NaCl in (A)
Elution: (B) 0% → (B) 75%, linear gradient
Flow rate: 0.86 mL/min (residence time 2 min)
Sample loaded: 1.5 mL

Transferrin (5 mg/mL)
BSA (10 mg/mL)
Pepsin (5 mg/mL)

Column: 6.6 mm I.D. x 50 mm
Equilibration buffer (A): 10 mM phosphate (pH 7)
Elution buffer (B): 1 M NaCl in (A)
Elution: (B) 0% → (B) 50%, linear gradient
Flow rate: 0.86 mL/min (residence time 2 min)
Sample loaded: 1.5 mL

Ribonuclease A (5 mg/mL)
Cytochrome C (2.5 mg/mL)
Lysozyme (1.5 mg/mL)

高耐盐性

Cellufine™ MAX IEX 层析介质在一系列盐浓度范围内保持高结合能力。该特性使得在使用Cellufine™ MAX IEX 层析介质之前省略了缓冲液交换步骤。

BSA DBC at 10% Breakthrough for Various NaCl Concentrations IgG DBC at 10% Breakthrough for Various NaCl Concentrations
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BSA concentration: 1 mg/mL
Column: 5 mm I.D. x 100 mm
Buffer: NaCl in 50 mM Tris-HCl (pH 8.5)
Flow rate: 300 cm/h (residence time 2 min)

IgG concentration: 1 mg/mL (polyclonal, human)
Column: 5 mm I.D. x 100 mm
Buffer: NaCl in 10 mM Na-acetate (pH 4.3)
Flow rate: 300 cm/h (residence time 2 min)

结合条件的优化

离子交换层析介质的结合能力取决于离子强度和pH值,分离条件需要根据目标蛋白质的特性进行优化。
下图显示了在所有测试的pH水平下,Cellufine™ MAX S-h 介质的IgG结合力高于含有10mM NaCl 的Agarose S 型介质。即使使用10mM NaCl,Cellufine™ MAX S-h在pH4.3下也具有较高的IgG DBC(动态结合能力)。

IgG DBC at 10% Breakthrough for Various Salt Concentrations and pH Levels

▶Cellufine™ MAX S-h ▶Agarose S Type Media
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IgG concentration: 1 mg/mL (polyclonal, human)
Column: 5 mm I.D. x 100 mm
Buffer: NaCl in 10 mM Na-acetate
Flow rate: 300 cm/h (residence time 2 min)

IgG concentration: 1 mg/mL (polyclonal, human)
Column: 5 mm I.D. x 100 mm
Buffer: NaCl in 10 mM Na-acetate
Flow rate: 300 cm/h (residence time 2 min)

特性和规格

Cellufine™ MAX IEX Series

ProductMAX Q-rMAX Q-hMAX S-rMAX S-hMAX DEAEMAX CM
Ligand Q Q S S DEAE CM
Ion exchange type Strong anion Strong cation Weak anion Weak cation
Support matrix Highly cross-linked cellulose with dextran scaffold
Particle size 40-130 µm
Ion exchange capacity 0.10-0.20 mEq/mL 0.13-0.22 mEq/mL 0.09-0.21 mEq/mL 0.10-0.22 mEq/mL 0.12-0.25 mEq/mL 0.09-0.22 mEq/mL
Adsorption capacityLysozyme - - ≥110 mg/mL ≥140 mg/mL - ≥140 mg/mL
BSA ≥120 mg/mL ≥190 mg/mL - - ≥120 mg/mL -
Chemical stability Stable in commonly used aqueous buffers and
cleaning solutions (NaOH up to 0.5 M)
pH stability 2-12 2-12 2-13 3-14 2-12 2-13
Operating pressure <0.3 MPa (3 bar, 44 psi)
Supplied as Suspension in 20% ethanol

Cellufine™ IEX Series

ProductQ-500A-200A-500A-800C-500
Ligand Q DEAE DEAE DEAE CM
Ion exchange type Strong anion Weak anion Weak cation
Support matrix Cross-linked cellulose
Particle size 40-130 µm
Degree of swelling 5-9
mL/g-dry wt
6-9
mL/g-dry wt
8-10
mL/g-dry wt
12-16
mL/g-dry wt
9-11
mL/g-dry wt
Ion exchange capacity 1.2-1.9
mEq/g-dry wt
0.8-1.1
mEq/g-dry wt
1.1-1.4
mEq/g-dry wt
0.6-1.0
mEq/g-dry wt
0.9-1.2
mEq/g-dry wt
Adsorption capacity Lysozyme - - - - ≥70 mg/mL
BSA ≥10 mg/mL ≥80 mg/mL ≥60 mg/mL ≥45 mg/mL -
Exclusion limit (protein) Approx.
500 kDa
Approx.
30 kDa
Approx.
500 kDa
Approx.
1,000 kDa
Approx.
500 kDa
Chemical stability Stable in commonly used aqueous buffers and
cleaning solutions (NaOH up to 0.5 M)
pH stability 1-13
Operating pressure <0.2 MPa (2 bar, 29 psi)
Supplied as Suspension in 20% ethanol

产品型号

► Cellufine™ MAX IEX Series

ProductQuantityCatalog Number
Cellufine™ MAX Q-r 1 mL x 5 (mini column) 20500-51
5 mL x 5 (mini column) 20500-55
100 mL 20500
500 mL 20501
5 L 20502
10 L 20503
Cellufine™ MAX Q-h 1 mL x 5 (mini column) 20600-51
5 mL x 5 (mini column) 20600-55
100 mL 20600
500 mL 20601
5 L 20602
10 L 20603
Cellufine™ MAX S-r 1 mL x 5 (mini column) 20300-51
5 mL x 5 (mini column) 20300-55
100 mL 20300
500 mL 20301
5 L 20302
10 L 20303
Cellufine™ MAX S-h 1 mL x 5 (mini column) 20400-51
5 mL x 5 (mini column) 20400-55
100 mL 20400
500 mL 20401
5 L 20402
10 L 20403
Cellufine™ MAX DEAE 1 mL x 5 (mini column) 21000-51
5 mL x 5 (mini column) 21000-55
100 mL 21000
500 mL 21001
5 L 21002
10 L 21003
Cellufine™ MAX CM 1 mL x 5 (mini column) 20900-51
5 mL x 5 (mini column) 20900-55
100 mL 20900
500 mL 20901
5 L 20902
10 L 20903

► Cellufine™ IEX Series

ProductQuantityCatalog Number
Cellufine™ Q-500 1 mL x 5 (mini column) 19907-51
5 mL x 5 (mini column) 19907-55
100 mL 675 982 327
500 mL 19907
5 L 19908
10 L 675 982 335
Cellufine™ A-200 1 mL x 5 (mini column) 19611-51
100 mL 676 980 327
500 mL 19611
5 L 19612
10 L 676 980 335
Cellufine™ A-500 1 mL x 5 (mini column) 19805-51
5 mL x 5 (mini column) 19805-55
100 mL 675 980 327
500 mL 19805
5 L 19806
10 L 675 980 335
Cellufine™ A-800 1 mL x 5 (mini column) 19865-51
5 mL x 5 (mini column) 19865-55
100 mL 673 980 327
500 mL 19800
5 L 19801
10 L 673 980 335
Cellufine™ C-500 1 mL x 5 (mini column) 19800-51
5 mL x 5 (mini column) 19800-55
100 mL 675 983 327
500 mL 19865
5 L 19866
10 L 675 983 335

接枝聚合物改性

MAX GS
用于单克隆抗体聚集体清除的离子交换层析介质

在单克隆抗体(mAb)生产工艺中(包括培养、蛋白A柱洗脱、低pH值病毒灭活和超滤/渗滤)聚集体形成是常见问题。这些mAb 聚集体可以降低药理活性并诱导过敏性免疫反应如过敏性休克,因此需要被清除。

Cellufine™ MAX GS 是一种强阳离子交换剂,用于清除mAb聚集体。

Cellufine™ MAX GS 基质由高度交联的纤维素组成。用含有离子交换基因的接枝链进行表面改性产生了一种专门的结构,为分离抗体单体和聚集体提供了独特的机制。孔直径也被优化以提供高吸附力。

Cellufine™ MAX GS 成为解决聚集体清除问题的宝贵工具。

Separation with Cellufine™ MAX GS vs. S agarose media for polyclonal IgG aggregates

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Poly IgG: thermal and acid stressed poly IgG
Buffer A: 10 mM acetate (pH 5.0) + 50 mM NaCl
Buffer B: 10 mM acetate (pH 5.0) + 1 M NaCl
Dynamic binding capacity of Cellufine™ MAX GS vs. S agarose media

优点

・有效分离抗体单体和聚集体
・高吸附力
・低压降
・与大规模柱层析兼容性极佳
・重复碱洗时的高稳定性
・简单的再生和去热源
・低非特异性吸附

Dynamic binding capacity of Cellufine™ MAX GS vs. S agarose media

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Column: 5mm ID x 50mm L
Polyclonal antibody: 1mg/mL in buffer A
Buffer A: 10mM acetate (pH 5.0) + 50 mM NaCl

Contour plot of DBC with Cellufine™ MAX S-h (upper) and Cellufine™ MAX GS (lower)

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Characteristics of Dynamic Binding Capacity (DBC) with Cellufine™ CEX

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▶Graft modified Cellufine™ MAX GS showed to be less susceptible to pH condition in comparison with dextran modified Cellufine™ MAX S-h.

Load condition

mAb: momoclonal antibody (5 mg/mL)
Actual bed volume: 0.59 mL (3 cm bed height)

▶Graft modified Cellufine™ MAX GS showed to be less susceptible to pH condition in comparison with dextran modified Cellufine™ MAX S-h.

使用Cellufine™ MAX GS 的MAb 聚集体分离
(低样品载量的梯度洗脱)

用梯度洗脱获得不同的MAb单体和二聚体峰值。

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Buffer A: 10 mM acetate (pH 4.5) + 50 mM NaCl

Buffer B: 10 mM acetate (pH 4.5) + 0.5 M NaCl

Gradient: 75 CV

Injection: acid-treated mAb1

产品型号

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Mixed mode resin, Cellufine™ MAX IB

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▶Benefit
Salt tolerance
Superior impurity removal
Less susceptible to buffer especially multivalent buffer
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亲和性

硫酸盐

Cellufine™ 硫酸盐层析介质适用于病毒,病菌/微生物抗原和肝素结合蛋白溶度,纯化和去热源的亲和介质。
与肝素层析不同,Cellufine™硫酸盐介质的配体不具有动源性,并且安全问题被最小化。

Structure of Cellufine™ Sulfate

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由非动源性亲和配体进行的病毒和蛋白结合

与传统的肝素层析相似,Cellufine™ 硫酸盐介质可用于各种病毒的纯化。在下面所示的流感病毒(H7N7)纯化研究中,实现了高浓度和高产率,而蛋白质和DNA被有效清除。
由于 Cellufine™ 硫酸盐介质与肝素层析不同,不使用动源性的配体,所以安全性问题被最小化。

Purification of Influenza Virus

Virus strain: Egg-derived A/duck/Hokkaido/vac-2/2004 (H7N7)
Column: Cellufine™ Sulfate, 70 mL (I.D. 22 mm x H 185 mm)
Flow rate: 100 cm/h
Equilibration buffer (A): 0.01 M phosphate (pH 7.4)
Washing buffer: 0.19 M NaCl in (A)
Elution buffer: 3 M NaCl in (A)

Notes:
1. HA-titer: Virus titer was estimated by hemagglutination assay.
2. A280 readings during parts of the load and wash stages of the run exceeded the detection limit and are shown as a dashed line.

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优越的化学稳定性

Cellufine™ 硫酸盐介质在酸性和碱性条件下比肝素琼脂糖层析介质更局稳定性。右图显示了Cellufine™ 硫酸盐介质即使在使用 0.2M HCl 和0.5M NaOH 的情况下也能在40天内保持其对溶菌酶的媳妇能力,而肝素琼脂糖介质在使用0.01M HCl 或0.5M NaOH的情况下吸附力急剧下降。

Chemical Stability of Cellufine™ Sulfate and Heparin Agarose Media

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高压操作下的线性流量

Cellufine™ 硫酸盐介质可以在高流量条件下运行。右图显示了Cellufine™ 硫酸盐介质的流速为线性,最高可达 250cm/h,即使在高压操作下,压缩的床高小于15%。
这些数据是通过使用三种不同批次Cellufine™ 硫酸盐介质的研究中获得的。

Flow Rate Curve of Cellufine™ Sulfate

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Column: 9 cm I.D. x 38 cm bed height
Mobile phase: pure water

特性和规格

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产品型号

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用于在下游加工过程中清除内毒素

Cellufine™ ET clean 是聚酯(ε-赖氨酸)固定化的Cellufine™(纤维素球形珠体)。珠体从样品溶液中结合并清除内毒素。聚酯(ε-赖氨酸)是由白色链霉菌产生的30-35个赖氨酸残基组成的微生物聚酯(氨基酸)。聚酯(ε-赖氨酸)配体和纤维素珠体是 JNC 公司的产品。

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The adsorption of endotoxin was measured as follows: The beads were washed and equilibrated with 0.02 M phosphate buffer (pH 7.0) – 0.05 M NaCl. The removal of endotoxin was determined using a batchwise method with 0.3 mL of wet beads and 2 mL of a protein solution (1 mg/mL) containing natural endotoxin. The suspension was shaken for 2 hr at 25 ℃. The endotoxin content was measured after filtrating through a 0.8 μm cellulose acetate filter to remove the beads from the suspension.

Molecular weight Exclusion

Cellufine™ ET Clean-S: 2 kDa
Cellufine™ ET Clean-L: 2000 kDa

特点
•由于稳定的偶联化学和纤维素基珠而产生的低配体泄漏。
•高内毒素负载量,
•耐盐性清除内毒素高达0.4M NaCl,
•由于其大孔径的蛋白质,ET clean-L与高MWt兼容。
•亲和配体是FDA批准的食品添加剂。
•1小时内蛋白样品中内毒素水平降低了>99%。
•简单的盒式注射器操作流程。
•可以放大直径达30cm 的柱子。

订购信息

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Cellufine™ 螯合物介质

Cellufine™ 螯合物介质设计用于蛋白质和肽的固定化的 金属螯合亲和层析。该介质包含固定亚氨基二乙酸(IDA)的交联纤维素。其刚性结构允许在高流速下操作,从而加快处理时间。当暴露于金属盐时,IDA快速与阳离子复合。所得到的金属螯合物部分(主要地)与表面可及残基如组氨酸标签相互所用。

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疏水作用

Hydrophobic Interaction Chromatography Media
Cellufine™ MAX Butyl,
Cellufine™ MAX Phenyl

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凝胶过滤

凝胶过滤层析介质
Cellufine™ GCL-2000HF

凝胶过滤或尺寸排阻层析已被证明是一种简单有效的蛋白质和其他生物分子纯化方法。然而,大多数凝胶过滤介质的传统缓慢流速和极限分辨力限制了它的实用性。这些填料的固有问题不仅存在于实验室制备规模上,而且还存在于工艺规模上。凝胶过滤的有限流量通常会造成严重的工艺瓶颈。. Cellufine™ GCL 介质克服了这些限值。球形纤维素基质的机械强度允许高流速,即使在大型工业柱中也是如此。

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分解范围
该介质提供了非常广泛的分解范围选择:从独特的 Cellufine™ GCL-2000 到用于超高分子量蛋白质复合物的Cellufine™ GCL-2000HF。下表显示了介质的排阻限值。

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该介质提供了非常广泛的分解范围选择:从独特的 Cellufine™ GCL-2000 到用于超高分子量蛋白质复合物的Cellufine™ GCL-2000HF。下表显示了介质的排阻限值。

凝胶过滤层析介质
Cellufine™ GH-25

用于快速蛋白质脱盐缓冲液交换和酒精及清洁剂的清除

Cellufine™ GH-25 脱盐介质是基于多孔、球形、高度交联的纤维素颗粒。尖锐的3kD排阻限值允许蛋白通过孔隙体积中的柱子,同时延缓内部孔隙中较小分子量的溶质。卓越的机械强度使其即使在大直径工艺规模的柱子中也可以在高流量下操作,从而将运行时间将至最低。

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应用
► 冷冻干燥或浓缩前脱盐
► 缓冲液交换
► 清除酒精或其他有机溶液
► 纯化核酸时清除芳香化合物(如苯酚)
► 去除用于溶解蛋白质的清洁剂(如 Triton X-100, SDS)
► 允许使用所有常用的溶剂和缓冲液而不收缩或膨胀
► 清除离液剂(如尿素、胍)

高速脱盐

虽然Cellufine™ GH-25的硬度使其非常适合大规模柱子的使用,但其在高流速下运行的能力使得在运行多个循环的较小柱子上使用获得与较大柱子相似的产量和较低的流速。与传统的层析不同,脱盐层析的性能(通过样品加载、脱盐和稀释进行测量)实际上可以随着流速的增加而提高,这是由于在高容量负载下样品稀释度降低。

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